CN103926375A - Device for simulating human digestive and glycolytic system and application method thereof - Google Patents

Device for simulating human digestive and glycolytic system and application method thereof Download PDF

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CN103926375A
CN103926375A CN201410089160.8A CN201410089160A CN103926375A CN 103926375 A CN103926375 A CN 103926375A CN 201410089160 A CN201410089160 A CN 201410089160A CN 103926375 A CN103926375 A CN 103926375A
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glycolysis
ware
digestion
chamber
large intestine
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CN103926375B (en
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聂少平
胡婕伦
谢明勇
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Nanchang University
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Nanchang University
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Abstract

The invention discloses a device for simulating a human digestive and glycolytic system and an application method thereof. The device comprises an oral digestion area, a gastric digestion area, a small intestine digestion area, a large intestine anterior-segment glycolysis area and a large intestine posterior-segment glycolysis area. An object to be measured is added into an inlet at the top of the device; all thermostatic waterbath system is turned on and the temperature remains constant at 37 DEG C; and all constant flow pumps and flow dividers are opened. Full dynamic flow of the object to be measured in the digestive system is realized, and the object to be measured successively undergoes oral digestion, gastric digestion, small intestine digestion, large intestine anterior-segment glycolysis and large intestine posterior-segment glycolysis. Digestion and glycolysis processes are simulated repeatedly for three times; digestion and glycolysis products at each part are collected; and digestion and glycolysis conditions of the object to be measured are detected. Simulation of digestion and glycolysis at each part is carried out completely according to the human digestive and glycolytic processes and physiological environment. Full dynamic simulation of digestive and glycolytic processes at each part is realized. Digestive and glycolytic conditions of various active or nutrient substances which enter human body can be observed. The device provided by the invention fits real human physiological environment.

Description

A kind of device and using method of simulating human body digestion glycolysis system
Technical field
The present invention relates to a kind of device and using method of simulating human body digestion glycolysis system.
Background technology
World's authoritative journal represents, research food or effect and the mechanism of action thereof of active substance in body are most important, because illustrating of mechanism of action can be for utilizing food or active substance that Research foundation is provided substantially in body, make it in human body, bring into play substantially physiological function, there is directive significance to avoiding it to produce possible adverse side effect simultaneously.The physiological function of the actual performance of food or active substance depends on what its product after human body digestion glycolysis is to a great extent.For the consideration of the moral ethics for studying in body, in-vitro simulated human body digestion glycolysis just becomes the key problem in technology of study on mechanism in food or active matter plastid.Key point by controlling human body digestion glycolysis process is carried out dynamic similation as temperature, digestive juice etc.Simultaneously, the digestion glycolysis system of simulation has good degree of accuracy compared with the digestion glycolysis system of human body, favorable reproducibility, easily control, can collect at any time digestion or the glycolysis product at each position, when experiment relates to the situation of toxic chemical substance, can not be subject to moral constraint, offer convenience to the research of the active substance mechanism of action.Therefore, in vitro digestion glycolysis is most important.
Different food or active nutritional material are not quite similar in the situation of human body digestion glycolysis, this makes the process of research digestion glycolysis very meaningful, research simultaneously knows that the digestion glycolysis of different material in human body can provide data for the whole digestion glycolysis case study of this material, simultaneously for the digestion glycolysis mode important in inhibiting of being familiar with and exploring different material.The Patents that relates in recent years digestion glycolysis simulation has: " intelligent biology body pipe intestinal digesting system simulation control device " (application number: 201220211927.6); " Simulative digestive device for gastrointestinal tract of animals " (application number: 200920105937.X); " Bionic digestive system for monogastric animals and the method based on this system simulation nonruminant digestion " (application number: 200910078147.1); " broiler external digestion simulating device (200810100818.5); " a kind of analog device of alimentary system " (application number: 200710078118.6).These patent reports have greatly enriched the device and the using method that are applicable to digest glycolysis, but up to the present, in these patents, for Mouthsimulator, concrete digestion does not describe in detail.Secondly, these patents do not have detailed simulation and explanation for the evacuation procedure of stomach in the simulation of stomach.These patents are also detailed not for the explanation of controlling parameter in small intestine simulated system and large intestine glycolysis system simultaneously.Wherein some patent is only basis with animal correlation parameter, little with people's relevance.And in this patent, undertaken by human body digestion glycolysis process and physiological environment completely for the digestion glycolysis simulation at each position first, each digestion glycolysis position is had been described in detail, simultaneously the process implementation at each digestion glycolysis position full dynamic similation, the real people's that fits physiological environment.
Summary of the invention
The object of the present invention is to provide a kind of device and using method of simulating human body digestion glycolysis system.The digestion glycolysis situation that this device and using method can enter after human body various activity or nutriment is observed, and helps to explore various activity or nutriment enters the digestion glycolysis situation after human body.
The present invention is achieved by the following technical solutions.
The device of simulation human body digestion glycolysis system of the present invention comprises that device comprises oral digestion district (A), peptic digest district (B), small intestine digestion zone (C), large intestine leading portion glycolysis district (D), large intestine back segment glycolysis district (E).Specifically comprise sample inlet (1), oral digestion chamber (2), peptic digest chamber (3), small intestine digestion chamber (4), large intestine leading portion glycolysis chamber (5), large intestine back segment glycolysis chamber (6), saliva ware (7), gastric juice ware (8), intestinal fluid ware (9), large intestine leading portion glycolysis nutrient culture media ware (10), large intestine back segment glycolysis nutrient culture media ware (11), the first dialysate chamber (12), the second dialysate chamber (13), airtight the first sieves (14), airtight the second sieves (15), oral digestion product ware (16), peptic digest product ware (17), small intestine digestion product ware (18), large intestine leading portion glycolysis product anaerobism ware (19), large intestine back segment glycolysis product anaerobism ware (20), residue glycolysis product reclaims ware (21).
Each parts annexation.
Oral digestion district (A): sample inlet (1) is connected with oral digestion chamber (2) right-hand member sample feeding pipe with diverting valve by connection valve, inside, the straight-through oral cavity digestion chamber (2) of oral digestion chamber (2) right-hand member sample feeding pipe.Oral digestion chamber (2) are all equipped with the water bath with thermostatic control circulation system in outside simultaneously.Oral digestion chamber (2) left end is connected with saliva ware (7) with a constant flow pump by flow diverter.Oral digestion chamber (2) left end outlet is by diverting valve, and connection valve is connected with oral digestion product ware (16) with constant flow pump.The middle part of oral digestion chamber (2) left end outlet is connected with peptic digest chamber (3) left end sample feeding pipe with connection valve by emptying delivery pipe.
Peptic digest district (B): inside, the straight-through peptic digest chamber (3) of peptic digest chamber (3) left end sample feeding pipe.Peptic digest chamber (3) is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with gastric juice ware (8) with a constant flow pump by flow diverter.Peptic digest chamber (3) right-hand member outlet is connected with a conduit by diverting valve, and this conduit other end is connected with peptic digest product ware (17) with constant flow pump by connection valve.Also be connected with small intestine digestion chamber (4) right-hand member sample feeding pipe by emptying delivery pipe in diverting valve junction.
Small intestine digestion zone (C): inside, the straight-through small intestine digestion chamber (4) of small intestine digestion chamber (4) right-hand member sample feeding pipe.Small intestine digestion chamber (4) is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with intestinal fluid ware (9) with a constant flow pump by flow diverter.Small intestine digestion chamber (4) left end outlet is connected with a conduit by diverting valve, and this conduit other end is connected with small intestine digestion product ware (18) with constant flow pump by connection valve.Also be connected with large intestine leading portion glycolysis chamber (5) left end sample feeding pipe by emptying delivery pipe in diverting valve junction.
Large intestine leading portion glycolysis district (D): inside, the straight-through large intestine leading portion glycolysis chamber (5) of large intestine leading portion glycolysis chamber (5) left end sample feeding pipe.Large intestine leading portion glycolysis chamber (5) is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with large intestine leading portion glycolysis nutrient culture media ware (10) with a constant flow pump by flow diverter.Large intestine leading portion glycolysis chamber (5) right-hand member outlet is connected with one end of airtight the first sieves (14) with conduit by diverting valve, airtight the first sieves (14) other end is connected with one end of the first dialysate chamber (12) by conduit, and the first dialysate chamber (12) other end is by a constant flow pump and large intestine leading portion glycolysis product anaerobism ware (19).Also be connected with large intestine back segment glycolysis chamber (6) right-hand member sample feeding pipe by emptying delivery pipe in diverting valve junction.
Large intestine back segment glycolysis district (E): inside, the straight-through large intestine back segment glycolysis chamber (6) of large intestine back segment glycolysis chamber (6) left end sample feeding pipe.Large intestine back segment glycolysis chamber (6) is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with large intestine back segment glycolysis nutrient culture media ware (11) with a constant flow pump by flow diverter.Large intestine back segment glycolysis chamber (5) left end outlet is connected with airtight the second sieves (15) one end with conduit by diverting valve, airtight the second sieves (15) other end is connected with the second dialysate chamber (13) one end by conduit, and the second dialysate chamber (13) other end is by a constant flow pump and large intestine back segment glycolysis product anaerobism ware (20).Also reclaiming ware (21) by emptying delivery pipe with residue glycolysis product in diverting valve junction is connected.
Using method: material to be digested is dissolved to (or being scattered in) with finite concentration make determinand solution or dispersion liquid in distilled water.To shift to an earlier date ready saliva, gastric juice, intestinal fluid, large intestine leading portion glycolysis nutrient culture media, large intestine back segment glycolysis nutrient culture media joins respectively in saliva ware (7), gastric juice ware (8), intestinal fluid ware (9), large intestine leading portion glycolysis nutrient culture media ware (10), large intestine back segment glycolysis nutrient culture media ware (11).Open whole water bath with thermostatic control systems and keep 37 DEG C of constant temperature.Open whole constant flow pumps and diverting valve.All the speed of constant flow pump is controlled at 4-6 ml/min.Determinand solution or dispersion liquid are added from the sample inlet (1) of the top of digestion glycolysis system and device, saliva joins oral digestion chamber (2) by constant flow pump and flow diverter from saliva ware (7), carry out oral digestion simulation, after oral digestion 10-15 min, the postdigestive collection of products of a part is in oral digestion product ware (16), and another part digestion product enters in the peptic digest chamber (3) in peptic digest district (B) with emptying delivery pipe.Gastric juice joins peptic digest chamber (3) by constant flow pump and flow diverter from gastric juice ware (8), carry out peptic digest simulation, after peptic digest 5-6 h, the postdigestive collection of products of a part is in peptic digest product ware (17), and another part digestion product enters in the small intestine digestion chamber (4) in small intestine digestion zone (C) with emptying delivery pipe.Intestinal fluid joins small intestine digestion chamber (4) by constant flow pump and flow diverter from intestinal fluid ware (9), after small intestine digestion 5-6 h, the postdigestive collection of products of a part is in small intestine digestion product ware (18), and another part digestion product enters in the large intestine leading portion glycolysis chamber (5) in large intestine leading portion glycolysis district (D) with emptying delivery pipe.Large intestine leading portion glycolysis nutrient culture media joins large intestine leading portion glycolysis chamber (5) by constant flow pump and flow diverter from large intestine leading portion glycolysis nutrient culture media ware (10), after large intestine leading portion glycolysis 10-12 h, product after part glycolysis is collected in large intestine leading portion glycolysis product anaerobism ware (19) by airtight the first sieves (14) and the first dialysate chamber (12), and another part glycolysis product enters in the large intestine back segment glycolysis chamber (6) in large intestine back segment glycolysis district (E) with emptying delivery pipe.Large intestine back segment glycolysis nutrient culture media joins large intestine back segment glycolysis chamber (6) by constant flow pump and flow diverter from large intestine back segment glycolysis nutrient culture media ware (11), after large intestine back segment glycolysis 10-12 h, product after part glycolysis is collected in large intestine back segment glycolysis product anaerobism ware (20) by airtight the second sieves (15) and the second dialysate chamber (13), and another part glycolysis product enters residue glycolysis product with emptying delivery pipe and reclaims in ware (21).Simulation digestion glycolysis process repeats 3 times.
Oral digestion product ware (16) is placed in boiling water bath (preparing in advance) a few minutes to remove the activity of ptyalin, detects afterwards the oral digestion situation of determinand in oral digestion product ware (16) with liquid phase method.Peptic digest product ware (17) is placed in boiling water bath (preparing in advance) a few minutes to remove pepsic activity, detects afterwards the peptic digest situation of determinand in peptic digest product ware (17) with liquid phase method.Small intestine digestion product ware (18) is placed in boiling water bath (preparing in advance) a few minutes to remove except pancreatin (comprising amylopsin and pancreatic lipase) and tryptic activity, detects afterwards the small intestine digestion situation of determinand in small intestine digestion product ware (18) with liquid phase method.Large intestine leading portion glycolysis product anaerobism ware (19) in boiling water bath (in advance prepare) 30-40 minute to remove activity of residue flora, and detect large intestine leading portion glycolysis product in large intestine leading portion glycolysis product anaerobism ware (19) with vapor phase method.Large intestine back segment glycolysis product anaerobism ware (20) in boiling water bath (in advance prepare) 30-40 minute to remove activity of residue flora, and detect large intestine back segment glycolysis product in large intestine back segment glycolysis product anaerobism ware (20) with vapor phase method.
The device of this simulation human body digestion glycolysis system and the craft feature of using method are that this device and using method digest glycolysis process by human body completely for the digestion glycolysis simulation at each position first and physiological environment is carried out, each digestion glycolysis position is had been described in detail, simultaneously the process implementation at each digestion glycolysis position full dynamic similation, the real people's that fits physiological environment.
The device of this simulation human body digestion glycolysis system and the technique effect of using method are: the digestion glycolysis situation that this device and using method can enter after human body various activity or nutriment is observed, and help to explore various activity or nutriment enters the digestion glycolysis situation after human body.
Brief description of the drawings
Fig. 1 is the device schematic diagram of simulation human body digestion glycolysis system.Wherein, A is oral digestion district; B is peptic digest district; C is small intestine digestion zone; D is large intestine leading portion glycolysis district; E is large intestine back segment glycolysis district.1 is sample inlet, 2 is oral digestion chamber, 3 is peptic digest chamber, 4 is small intestine digestion chamber, 5 is large intestine leading portion glycolysis chamber, 6 is large intestine back segment glycolysis chamber, 7 is saliva ware, 8 is gastric juice ware, 9 is intestinal fluid ware, 10 is large intestine leading portion glycolysis nutrient culture media ware, 11 is large intestine back segment glycolysis nutrient culture media ware, 12 is the first dialysate chamber, 13 is the second dialysate chamber, 14 is the first airtight sieves, 15 is the second airtight sieves, 16 is oral digestion product ware, 17 is peptic digest product ware, 18 is small intestine digestion product ware, 19 is large intestine leading portion glycolysis product anaerobism ware, 20 is large intestine back segment glycolysis product anaerobism ware, 21 is residue glycolysis product recovery ware.
Fig. 2 is F-F the first half enlarged drawing in the device schematic diagram of simulation human body digestion glycolysis system of Fig. 1.
Fig. 3 is F-F the latter half enlarged drawing in the device schematic diagram of simulation human body digestion glycolysis system of Fig. 1.
Embodiment
Embodiment 1.
The human body digestion glycolysis simulation of plant polyose.
1. raw material is prepared.
(1) saliva.
Fresh clarification saliva sample is collected from a healthy volunteer.This volunteer does not have chronic disease and within least 3 months, does not use the medicine of antibiotics.Before saliva collection, this volunteer is limited diet and drinks water.Collect the mode that adopts direct ptysis.By the saliva of having collected, under 1500 x g speed, centrifugal 10 min are to remove cell, and supernatant is preserved stand-by at-20 DEG C.
(2) gastric juice.
Gastric juice is to take 30 mg gastric lipases, and 30 mg pepsins join the (preparation of stomach electrolyte solution: take 3 g NaCl, 0.5 g KCl, 0.1 g CaCl in 150 g stomach electrolyte 2, 0.5 g NaHCO 3, in the volumetric flask of 1 L, electrolyte solution pH is adjusted to 3 with the HCl of 0.1 M with deionized water constant volume), then add 1 mL CH 3cOONa(1 M, pH 5).Room temperature lower magnetic force stirs 10 min, pH value of solution is adjusted to 3 with the HCl of 0.1 M, is placed in refrigerator for subsequent use.
(3) intestinal fluid.
Intestinal fluid is to take respectively (the intestines electrolyte solution preparation: take 5 g NaCl, 0.5 g KCl, 0.3 g CaCl of 100 g intestines electrolyte solutions 2, be surely dissolved in the volumetric flask of 1 L with deionized water, pH value of solution is adjusted to 7 with the NaOH of 0.1 M), 100 g(7%, w/w) pancreatin solution, the trypsase of 12 mg, 180 g water.Mixed solution pH is adjusted to 7 with the NaOH of 0.1 M.Add bile powder (2%, w/v) in solution, to make intestinal fluid simultaneously.
(4) large intestine leading portion glycolysis nutrient culture media.
In 1 L glycolysis nutrient culture media, contain: 4 g NaCl, 4 g KCl, 1 g NaHCO 3, 0.6 g MgSO 4h 2o, 0.5 g KH 2pO 4, 0.5 g K 2hPO 4, 0.05 g CaCl 2, 0.005 g FeSO 47H 2o, 1 ml Tween 80 and the reddish black solution of 3 ml resin (0.025%, w/v, anaerobism indicator).Nutrient culture media is stand-by after sterilizing 15 min at 121 DEG C.In glycolysis nutrient culture media, add afterwards 150 g human excrement and urines (volunteer of ight soil takes in normal diet under normal circumstances, there is no digestive disease, does not take microbiotic at least 3 months).
(5) large intestine back segment glycolysis nutrient culture media.
In 1 L glycolysis nutrient culture media, contain: 4 g NaCl, 4 g KCl, 1 g NaHCO 3, 0.6 g MgSO 4h 2o, 0.7 g Cys HClH 2o, 0.5 g KH 2pO 4, 0.5 g K 2hPO 4, 0.4 g bile salt, 0.05 g CaCl 2, 0.005 g FeSO 47H 2o, 1 ml Tween 80 and the reddish black solution of 3 ml resin (0.025%, w/v, anaerobism indicator).Nutrient culture media is stand-by after sterilizing 15 min at 121 DEG C.In glycolysis nutrient culture media, add afterwards 250 g human excrement and urines (volunteer of ight soil takes in normal diet under normal circumstances, there is no digestive disease, does not take microbiotic at least 3 months).
2. simulation human body digestion glycolysis systematic procedure.
The device schematic diagram of simulation human body digestion glycolysis system as shown in Figure 1.Device comprises oral digestion district A, peptic digest district B, small intestine digestion zone C, large intestine leading portion glycolysis district D, large intestine back segment glycolysis district E.Specifically be included as sample inlet 1, oral digestion chamber 2, peptic digest chamber 3, small intestine digestion chamber 4, large intestine leading portion glycolysis chamber 5, large intestine back segment glycolysis chamber 6, saliva ware 7, gastric juice ware 8, intestinal fluid ware 9, large intestine leading portion glycolysis nutrient culture media ware 10, large intestine back segment glycolysis nutrient culture media ware 11, it is the first dialysate chamber 12, the second dialysate chamber 13, the first airtight sieves 14, the second airtight sieves 15, oral digestion product ware 16, peptic digest product ware 17, small intestine digestion product ware 18, large intestine leading portion glycolysis product anaerobism ware 19, large intestine back segment glycolysis product anaerobism ware 20, residue glycolysis product reclaims ware 21.
Each parts annexation.
Oral digestion district A: sample inlet 1 is connected with oral digestion chamber 2 right-hand member sample feeding pipes with diverting valve by connection valve, 2 inside, digestion chamber, the straight-through oral cavity of oral digestion chamber 2 right-hand member sample feeding pipes.The water bath with thermostatic control circulation system is all equipped with in outside in oral digestion chamber 2 simultaneously.Oral digestion chamber 2 left ends are connected with saliva ware 7 with a constant flow pump by flow diverter.Oral digestion chamber 2 left end outlets are by diverting valve, and connection valve is connected with oral digestion product ware 16 with constant flow pump.The middle part of oral digestion chamber 2 left end outlets is connected with peptic digest chamber 3 left end sample feeding pipes with connection valve by emptying delivery pipe.
Peptic digest district B: 3 inside, the straight-through peptic digest chamber of peptic digest chamber 3 left end sample feeding pipes.Peptic digest chamber 3 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with gastric juice ware 8 with a constant flow pump by flow diverter.Peptic digest chamber 3 right-hand member outlets are connected with a conduit by diverting valve, and this conduit other end is connected with peptic digest product ware 17 with constant flow pump by connection valve.Also be connected with small intestine digestion chamber 4 right-hand member sample feeding pipes by emptying delivery pipe in diverting valve junction.
Small intestine digestion zone C: 4 inside, the straight-through small intestine digestion chamber of small intestine digestion chamber 4 right-hand member sample feeding pipes.Small intestine digestion chamber 4 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with intestinal fluid ware 9 with a constant flow pump by flow diverter.Small intestine digestion chamber 4 left end outlets are connected with a conduit by diverting valve, and this conduit other end is connected with small intestine digestion product ware 18 with constant flow pump by connection valve.Also be connected with large intestine leading portion glycolysis chamber 5 left end sample feeding pipes by emptying delivery pipe in diverting valve junction.
Large intestine leading portion glycolysis district D: 5 inside, the straight-through large intestine leading portion glycolysis chamber of large intestine leading portion glycolysis chamber 5 left end sample feeding pipes.Large intestine leading portion glycolysis chamber 5 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with large intestine leading portion glycolysis nutrient culture media ware 10 with a constant flow pump by flow diverter.Large intestine leading portion glycolysis chamber 5 right-hand member outlets are connected with one end of airtight the first sieves 14 by diverting valve and conduit, first airtight sieves 14 other ends are connected with one end of the first dialysate chamber 12 by conduit, and first dialysate chamber 12 other ends are by a constant flow pump and large intestine leading portion glycolysis product anaerobism ware 19.Also be connected with large intestine back segment glycolysis chamber 6 right-hand member sample feeding pipes by emptying delivery pipe in diverting valve junction.
Large intestine back segment glycolysis district E: 6 inside, the straight-through large intestine back segment glycolysis chamber of large intestine back segment glycolysis chamber 6 left end sample feeding pipes.Large intestine back segment glycolysis chamber 6 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with large intestine back segment glycolysis nutrient culture media ware 11 with a constant flow pump by flow diverter.Large intestine back segment glycolysis chamber 5 left end outlets are connected with second airtight sieves 15 one end with conduit by diverting valve, second airtight sieves 15 other ends are connected with second dialysate chamber 13 one end by conduit, and second dialysate chamber 13 other ends are by a constant flow pump and large intestine back segment glycolysis product anaerobism ware 20.Also reclaiming ware 21 by emptying delivery pipe with residue glycolysis product in diverting valve junction is connected.
Using method: plant polyose solution is made into certain concentration solution (5 mg/ml).To shift to an earlier date ready saliva, gastric juice, intestinal fluid, large intestine leading portion glycolysis nutrient culture media, large intestine back segment glycolysis nutrient culture media joins respectively in saliva ware 7, gastric juice ware 8, intestinal fluid ware 9, large intestine leading portion glycolysis nutrient culture media ware 10, large intestine back segment glycolysis nutrient culture media ware 11.Open whole water bath with thermostatic control systems and keep 37 DEG C of constant temperature.Open whole constant flow pumps and diverting valve.All the speed of constant flow pump is controlled at 5 ml/min.Plant polyose solution is added from the sample inlet 1 of the top of digestion glycolysis system and device, saliva joins oral digestion chamber 2 by constant flow pump and flow diverter from saliva ware 7, carry out oral digestion simulation, after oral digestion 10 min, the postdigestive collection of products of a part obtains plant polyose oral digestion product in oral digestion product ware 16, and another part digestion product enters in the peptic digest chamber 3 in peptic digest district B with emptying delivery pipe.Gastric juice joins peptic digest chamber 3 by constant flow pump and flow diverter from gastric juice ware 8, carry out peptic digest simulation, after peptic digest 6 h, the postdigestive collection of products of a part obtains plant polyose peptic digest product in peptic digest product ware 17, and another part digestion product enters in the small intestine digestion chamber 4 in the C of small intestine digestion zone with emptying delivery pipe.Intestinal fluid joins small intestine digestion chamber 4 by constant flow pump and flow diverter from intestinal fluid ware 9, small intestine digests after 6 h, the postdigestive collection of products of a part obtains plant polyose small intestine digestion product in small intestine digestion product ware 18, and another part digestion product enters in the large intestine leading portion glycolysis chamber 5 in large intestine leading portion glycolysis district D with emptying delivery pipe.Large intestine leading portion glycolysis nutrient culture media joins large intestine leading portion glycolysis chamber 5 by constant flow pump and flow diverter from large intestine leading portion glycolysis nutrient culture media ware 10, after large intestine leading portion glycolysis 12 h, product after part glycolysis is collected in large intestine leading portion glycolysis product anaerobism ware 19 and is obtained plant polyose large intestine leading portion glycolysis product by the first airtight sieves 14 and the first dialysate chamber 12, and another part glycolysis product enters in the large intestine back segment glycolysis chamber 6 in large intestine back segment glycolysis district E with emptying delivery pipe.Large intestine back segment glycolysis nutrient culture media joins large intestine back segment glycolysis chamber 6 by constant flow pump and flow diverter from large intestine back segment glycolysis nutrient culture media ware 11, after large intestine back segment glycolysis 12 h, product after part glycolysis is collected in large intestine back segment glycolysis product anaerobism ware 20 and is obtained plant polyose large intestine back segment glycolysis product by the second airtight sieves 15 and the second dialysate chamber 13, and another part glycolysis product enters residue glycolysis product with emptying delivery pipe 15 and reclaims in ware 21.Simulation digestion glycolysis process repeats 3 times.
Oral digestion product ware 16 is placed in 5 minutes activity with removal ptyalin of boiling water bath (preparing in advance), detects afterwards the oral digestion situation (the change of molecular weight) of plant polyose in oral digestion product ware 16 with liquid phase method.Peptic digest product ware 17 is placed in boiling water bath (in advance prepare) 5 minutes to remove pepsic activity, detect afterwards the peptic digest situation (the change of molecular weight) of plant polyose in peptic digest product ware 17 with liquid phase method, and with the quantitative change of dinitrosalicylic acid method detection reducing sugar.Small intestine digestion product ware 18 is placed in boiling water bath (preparing in advance) and removes pancreatin (comprising amylopsin and pancreatic lipase) and tryptic activity to remove in 5 minutes, detect afterwards the small intestine digestion situation (the change of molecular weight) of plant polyose in small intestine digestion product ware 18 with liquid phase method, and with the quantitative change of dinitrosalicylic acid method detection reducing sugar.Large intestine leading portion glycolysis product anaerobism ware 19 (is prepared in advance) 40 minutes to remove the activity of residue flora in boiling water bath, and detects the large intestine leading portion glycolysis product of plant polyose in large intestine leading portion glycolysis product anaerobism ware 19 with vapor phase method.Large intestine back segment glycolysis product anaerobism ware 20 (is prepared in advance) 40 minutes to remove the activity of residue flora in boiling water bath, and detects the large intestine back segment glycolysis product of plant polyose in large intestine back segment glycolysis product anaerobism ware 20 with vapor phase method.
Testing result: this plant polyose can not be decomposed in human oral cavity digestion.After stomach simulation digestion, molecular weight significantly reduces, and reducing sugar amount significantly raises, and shows that this plant polyose is decomposed in human stomach's digestion, and part glycosidic bond is interrupted simultaneously.After small intestine simulation digestion, molecular weight reduces on a small quantity subsequently, and reducing sugar amount has certain rising, shows that this plant polyose is decomposed on a small quantity in the digestion of human small intestine's liquid, and simultaneously a small amount of glycosidic bond is interrupted.Through the glycolysis of large intestine leading portion and after the glycolysis of large intestine back segment, can produce a large amount of SCFAs, significantly increase acetic acid, the amount of propionic acid and butyric acid, this will be conducive to the large gut health of human body.
Embodiment 2.
The human body digestion glycolysis simulation of cornstarch.
1. raw material is prepared.
(1) saliva.
Fresh clarification saliva sample is collected from a healthy volunteer.This volunteer does not have chronic disease and within least 3 months, does not use the medicine of antibiotics.Before saliva collection, this volunteer is limited diet and drinks water.Collect the mode that adopts direct ptysis.By the saliva of having collected, under 1500 x g speed, centrifugal 10 min are to remove cell, and supernatant is preserved stand-by at-20 DEG C.
(2) gastric juice.
Gastric juice is to take 30 mg gastric lipases, and 30 mg pepsins join the (preparation of stomach electrolyte solution: take 3 g NaCl, 0.5 g KCl, 0.1 g CaCl in 150 g stomach electrolyte 2, 0.5 g NaHCO 3, in the volumetric flask of 1 L, electrolyte solution pH is adjusted to 3 with the HCl of 0.1 M with deionized water constant volume), then add 1 mL CH 3cOONa(1 M, pH 5).Room temperature lower magnetic force stirs 10 min, pH value of solution is adjusted to 3 with the HCl of 0.1 M, is placed in refrigerator for subsequent use.
(3) intestinal fluid.
Intestinal fluid is to take respectively (the intestines electrolyte solution preparation: take 5 g NaCl, 0.5 g KCl, 0.3 g CaCl of 100 g intestines electrolyte solutions 2, be surely dissolved in the volumetric flask of 1 L with deionized water, pH value of solution is adjusted to 7 with the NaOH of 0.1 M), 100 g(7%, w/w) pancreatin solution, the trypsase of 12 mg, 180 g water.Mixed solution pH is adjusted to 7 with the NaOH of 0.1 M.Add bile powder (2%, w/v) in solution, to make intestinal fluid simultaneously.
(4) large intestine leading portion glycolysis nutrient culture media.
In 1 L glycolysis nutrient culture media, contain: 4 g NaCl, 4 g KCl, 1 g NaHCO 3, 0.6 g MgSO 4h 2o, 0.5 g KH 2pO 4, 0.5 g K 2hPO 4, 0.05 g CaCl 2, 0.005 g FeSO 47H 2o, 1 ml Tween 80 and the reddish black solution of 3 ml resin (0.025%, w/v, anaerobism indicator).Nutrient culture media is stand-by after sterilizing 15 min at 121 DEG C.In glycolysis nutrient culture media, add afterwards 150 g human excrement and urines (volunteer of ight soil takes in normal diet under normal circumstances, there is no digestive disease, does not take microbiotic at least 3 months).
(5) large intestine back segment glycolysis nutrient culture media.
In 1 L glycolysis nutrient culture media, contain: 4 g NaCl, 4 g KCl, 1 g NaHCO 3, 0.6 g MgSO 4h 2o, 0.7 g Cys HClH 2o, 0.5 g KH 2pO 4, 0.5 g K 2hPO 4, 0.4 g bile salt, 0.05 g CaCl 2, 0.005 g FeSO 47H 2o, 1 ml Tween 80 and the reddish black solution of 3 ml resin (0.025%, w/v, anaerobism indicator).Nutrient culture media is stand-by after sterilizing 15 min at 121 DEG C.In glycolysis nutrient culture media, add afterwards 250 g human excrement and urines (volunteer of ight soil takes in normal diet under normal circumstances, there is no digestive disease, does not take microbiotic at least 3 months).
2. simulation human body digestion glycolysis systematic procedure.
The device schematic diagram of simulation human body digestion glycolysis system as shown in Figure 1.Device comprises oral digestion district A, peptic digest district B, small intestine digestion zone C, large intestine leading portion glycolysis district D, large intestine back segment glycolysis district E.Specifically be included as sample inlet 1, oral digestion chamber 2, peptic digest chamber 3, small intestine digestion chamber 4, large intestine leading portion glycolysis chamber 5, large intestine back segment glycolysis chamber 6, saliva ware 7, gastric juice ware 8, intestinal fluid ware 9, large intestine leading portion glycolysis nutrient culture media ware 10, large intestine back segment glycolysis nutrient culture media ware 11, it is the first dialysate chamber 12, the second dialysate chamber 13, the first airtight sieves 14, the second airtight sieves 15, oral digestion product ware 16, peptic digest product ware 17, small intestine digestion product ware 18, large intestine leading portion glycolysis product anaerobism ware 19, large intestine back segment glycolysis product anaerobism ware 20, residue glycolysis product reclaims ware 21.
Each parts annexation.
Oral digestion district A: sample inlet 1 is connected with oral digestion chamber 2 right-hand member sample feeding pipes with diverting valve by connection valve, 2 inside, digestion chamber, the straight-through oral cavity of oral digestion chamber 2 right-hand member sample feeding pipes.The water bath with thermostatic control circulation system is all equipped with in outside in oral digestion chamber 2 simultaneously.Oral digestion chamber 2 left ends are connected with saliva ware 7 with a constant flow pump by flow diverter.Oral digestion chamber 2 left end outlets are by diverting valve, and connection valve is connected with oral digestion product ware 16 with constant flow pump.The middle part of oral digestion chamber 2 left end outlets is connected with peptic digest chamber 3 left end sample feeding pipes with connection valve by emptying delivery pipe.
Peptic digest district B: 3 inside, the straight-through peptic digest chamber of peptic digest chamber 3 left end sample feeding pipes.Peptic digest chamber 3 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with gastric juice ware 8 with a constant flow pump by flow diverter.Peptic digest chamber 3 right-hand member outlets are connected with a conduit by diverting valve, and this conduit other end is connected with peptic digest product ware 17 with constant flow pump by connection valve.Also be connected with small intestine digestion chamber 4 right-hand member sample feeding pipes by emptying delivery pipe in diverting valve junction.
Small intestine digestion zone C: 4 inside, the straight-through small intestine digestion chamber of small intestine digestion chamber 4 right-hand member sample feeding pipes.Small intestine digestion chamber 4 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with intestinal fluid ware 9 with a constant flow pump by flow diverter.Small intestine digestion chamber 4 left end outlets are connected with a conduit by diverting valve, and this conduit other end is connected with small intestine digestion product ware 18 with constant flow pump by connection valve.Also be connected with large intestine leading portion glycolysis chamber 5 left end sample feeding pipes by emptying delivery pipe in diverting valve junction.
Large intestine leading portion glycolysis district D: 5 inside, the straight-through large intestine leading portion glycolysis chamber of large intestine leading portion glycolysis chamber 5 left end sample feeding pipes.Large intestine leading portion glycolysis chamber 5 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with large intestine leading portion glycolysis nutrient culture media ware 10 with a constant flow pump by flow diverter.Large intestine leading portion glycolysis chamber 5 right-hand member outlets are connected with one end of airtight the first sieves 14 by diverting valve and conduit, first airtight sieves 14 other ends are connected with one end of the first dialysate chamber 12 by conduit, and first dialysate chamber 12 other ends are by a constant flow pump and large intestine leading portion glycolysis product anaerobism ware 19.Also be connected with large intestine back segment glycolysis chamber 6 right-hand member sample feeding pipes by emptying delivery pipe in diverting valve junction.
Large intestine back segment glycolysis district E: 6 inside, the straight-through large intestine back segment glycolysis chamber of large intestine back segment glycolysis chamber 6 left end sample feeding pipes.Large intestine back segment glycolysis chamber 6 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with large intestine back segment glycolysis nutrient culture media ware 11 with a constant flow pump by flow diverter.Large intestine back segment glycolysis chamber 5 left end outlets are connected with second airtight sieves 15 one end with conduit by diverting valve, second airtight sieves 15 other ends are connected with second dialysate chamber 13 one end by conduit, and second dialysate chamber 13 other ends are by a constant flow pump and large intestine back segment glycolysis product anaerobism ware 20.Also reclaiming ware 21 by emptying delivery pipe with residue glycolysis product in diverting valve junction is connected.
Using method: corn starch solution is made into certain concentration solution (5 mg/ml).To shift to an earlier date ready saliva, gastric juice, intestinal fluid, large intestine leading portion glycolysis nutrient culture media, large intestine back segment glycolysis nutrient culture media joins respectively in saliva ware 7, gastric juice ware 8, intestinal fluid ware 9, large intestine leading portion glycolysis nutrient culture media ware 10, large intestine back segment glycolysis nutrient culture media ware 11.Open whole water bath with thermostatic control systems and keep 37 DEG C of constant temperature.Open whole constant flow pumps and diverting valve.All the speed of constant flow pump is controlled at 6 ml/min.Corn starch solution is added from the sample inlet 1 of the top of digestion glycolysis system and device, saliva joins oral digestion chamber 2 by constant flow pump and flow diverter from saliva ware 7, carry out oral digestion simulation, after oral digestion 15 min, the postdigestive collection of products of a part obtains cornstarch oral digestion product in oral digestion product ware 16, and another part digestion product enters in the peptic digest chamber 3 in peptic digest district B with emptying delivery pipe.Gastric juice joins peptic digest chamber 3 by constant flow pump and flow diverter from gastric juice ware 8, carry out peptic digest simulation, after peptic digest 6 h, the postdigestive collection of products of a part obtains cornstarch peptic digest product in peptic digest product ware 17, and another part digestion product enters in the small intestine digestion chamber 4 in the C of small intestine digestion zone with emptying delivery pipe.Intestinal fluid joins small intestine digestion chamber 4 by constant flow pump and flow diverter from intestinal fluid ware 9, small intestine digests after 6 h, the postdigestive collection of products of a part obtains cornstarch small intestine digestion product in small intestine digestion product ware 18, and another part digestion product enters in the large intestine leading portion glycolysis chamber 5 in large intestine leading portion glycolysis district D with emptying delivery pipe.Large intestine leading portion glycolysis nutrient culture media joins large intestine leading portion glycolysis chamber 5 by constant flow pump and flow diverter from large intestine leading portion glycolysis nutrient culture media ware 10, after large intestine leading portion glycolysis 12 h, product after part glycolysis is collected in large intestine leading portion glycolysis product anaerobism ware 19 and is obtained cornstarch large intestine leading portion glycolysis product by the first airtight sieves 14 and the first dialysate chamber 12, and another part glycolysis product enters in the large intestine back segment glycolysis chamber 6 in large intestine back segment glycolysis district E with emptying delivery pipe.Large intestine back segment glycolysis nutrient culture media joins large intestine back segment glycolysis chamber 6 by constant flow pump and flow diverter from large intestine back segment glycolysis nutrient culture media ware 11, after large intestine back segment glycolysis 12 h, product after part glycolysis is collected in large intestine back segment glycolysis product anaerobism ware 20 and is obtained cornstarch large intestine back segment glycolysis product by the second airtight sieves 15 and the second dialysate chamber 13, and another part glycolysis product enters residue glycolysis product with emptying delivery pipe 15 and reclaims in ware 21.Simulation digestion glycolysis process repeats 3 times.
Oral digestion product ware 16 is placed in 5 minutes activity with removal ptyalin of boiling water bath (preparing in advance), detects afterwards the oral digestion situation (the change of molecular weight) of cornstarch in oral digestion product ware 16 with liquid phase method.Peptic digest product ware 17 is placed in boiling water bath (in advance prepare) 5 minutes to remove pepsic activity, detect afterwards the peptic digest situation (the change of molecular weight) of cornstarch in peptic digest product ware 17 with liquid phase method, and with the quantitative change of dinitrosalicylic acid method detection reducing sugar.Small intestine digestion product ware 18 is placed in boiling water bath (preparing in advance) and removes pancreatin (comprising amylopsin and pancreatic lipase) and tryptic activity to remove in 5 minutes, detect afterwards the small intestine digestion situation (the change of molecular weight) of cornstarch in small intestine digestion product ware 18 with liquid phase method, and with the quantitative change of dinitrosalicylic acid method detection reducing sugar.Large intestine leading portion glycolysis product anaerobism ware 19 (is prepared in advance) 30 minutes to remove the activity of residue flora in boiling water bath, and detects the large intestine leading portion glycolysis product of cornstarch in large intestine leading portion glycolysis product anaerobism ware 19 with vapor phase method.Large intestine back segment glycolysis product anaerobism ware 20 (is prepared in advance) 30 minutes to remove the activity of residue flora in boiling water bath, and detects the large intestine back segment glycolysis product of cornstarch in large intestine back segment glycolysis product anaerobism ware 20 with vapor phase method.
Testing result: this cornstarch is after the simulation digestion of oral cavity, and molecular weight significantly reduces, and reducing sugar amount significantly raises, shows that this cornstarch is decomposed in human oral cavity digestion, and part glycosidic bond is interrupted simultaneously.After stomach simulation digestion, molecular weight part reduces, and reducing sugar amount has certain rising, shows that this cornstarch is decomposed on a small quantity in human stomach's digestion, and simultaneously a small amount of glycosidic bond is interrupted.After small intestine simulation digestion, molecular weight significantly reduces subsequently, and reducing sugar amount significantly raises, and shows that this cornstarch is decomposed in a large number in human small intestine's digestion, and simultaneously a large amount of glycosidic bonds are interrupted.Finally by crossing after the glycolysis of large intestine leading portion and back segment, can produce a large amount of SCFAs, significantly increase acetic acid, the amount of propionic acid and butyric acid.
Embodiment 3.
The human body digestion glycolysis simulation of soybean protein.
1. raw material is prepared.
(1) saliva.
Fresh clarification saliva sample is collected from a healthy volunteer.This volunteer does not have chronic disease and within least 3 months, does not use the medicine of antibiotics.Before saliva collection, this volunteer is limited diet and drinks water.Collect the mode that adopts direct ptysis.By the saliva of having collected, under 1500 x g speed, centrifugal 10 min are to remove cell, and supernatant is preserved stand-by at-20 DEG C.
(2) gastric juice.
Gastric juice is to take 30 mg gastric lipases, and 30 mg pepsins join the (preparation of stomach electrolyte solution: take 3 g NaCl, 0.5 g KCl, 0.1 g CaCl in 150 g stomach electrolyte 2, 0.5 g NaHCO 3, in the volumetric flask of 1 L, electrolyte solution pH is adjusted to 3 with the HCl of 0.1 M with deionized water constant volume), then add 1 mL CH 3cOONa(1 M, pH 5).Room temperature lower magnetic force stirs 10 min, pH value of solution is adjusted to 3 with the HCl of 0.1 M, is placed in refrigerator for subsequent use.
(3) intestinal fluid.
Intestinal fluid is to take respectively (the intestines electrolyte solution preparation: take 5 g NaCl, 0.5 g KCl, 0.3 g CaCl of 100 g intestines electrolyte solutions 2, be surely dissolved in the volumetric flask of 1 L with deionized water, pH value of solution is adjusted to 7 with the NaOH of 0.1 M), 100 g(7%, w/w) pancreatin solution, the trypsase of 12 mg, 180 g water.Mixed solution pH is adjusted to 7 with the NaOH of 0.1 M.Add bile powder (2%, w/v) in solution, to make intestinal fluid simultaneously.
(4) large intestine leading portion glycolysis nutrient culture media.
In 1 L glycolysis nutrient culture media, contain: 4 g NaCl, 4 g KCl, 1 g NaHCO 3, 0.6 g MgSO 4h 2o, 0.5 g KH 2pO 4, 0.5 g K 2hPO 4, 0.05 g CaCl 2, 0.005 g FeSO 47H 2o, 1 ml Tween 80 and the reddish black solution of 3 ml resin (0.025%, w/v, anaerobism indicator).Nutrient culture media is stand-by after sterilizing 15 min at 121 DEG C.In glycolysis nutrient culture media, add afterwards 150 g human excrement and urines (volunteer of ight soil takes in normal diet under normal circumstances, there is no digestive disease, does not take microbiotic at least 3 months).
(5) large intestine back segment glycolysis nutrient culture media.
In 1 L glycolysis nutrient culture media, contain: 4 g NaCl, 4 g KCl, 1 g NaHCO 3, 0.6 g MgSO 4h 2o, 0.7 g Cys HClH 2o, 0.5 g KH 2pO 4, 0.5 g K 2hPO 4, 0.4 g bile salt, 0.05 g CaCl 2, 0.005 g FeSO 47H 2o, 1 ml Tween 80 and the reddish black solution of 3 ml resin (0.025%, w/v, anaerobism indicator).Nutrient culture media is stand-by after sterilizing 15 min at 121 DEG C.In glycolysis nutrient culture media, add afterwards 250 g human excrement and urines (volunteer of ight soil takes in normal diet under normal circumstances, there is no digestive disease, does not take microbiotic at least 3 months).
2. simulation human body digestion glycolysis systematic procedure.
The device schematic diagram of simulation human body digestion glycolysis system as shown in Figure 1.Device comprises oral digestion district A, peptic digest district B, small intestine digestion zone C, large intestine leading portion glycolysis district D, large intestine back segment glycolysis district E.Specifically be included as sample inlet 1, oral digestion chamber 2, peptic digest chamber 3, small intestine digestion chamber 4, large intestine leading portion glycolysis chamber 5, large intestine back segment glycolysis chamber 6, saliva ware 7, gastric juice ware 8, intestinal fluid ware 9, large intestine leading portion glycolysis nutrient culture media ware 10, large intestine back segment glycolysis nutrient culture media ware 11, it is the first dialysate chamber 12, the second dialysate chamber 13, the first airtight sieves 14, the second airtight sieves 15, oral digestion product ware 16, peptic digest product ware 17, small intestine digestion product ware 18, large intestine leading portion glycolysis product anaerobism ware 19, large intestine back segment glycolysis product anaerobism ware 20, residue glycolysis product reclaims ware 21.
Each parts annexation.
Oral digestion district A: sample inlet 1 is connected with oral digestion chamber 2 right-hand member sample feeding pipes with diverting valve by connection valve, 2 inside, digestion chamber, the straight-through oral cavity of oral digestion chamber 2 right-hand member sample feeding pipes.The water bath with thermostatic control circulation system is all equipped with in outside in oral digestion chamber 2 simultaneously.Oral digestion chamber 2 left ends are connected with saliva ware 7 with a constant flow pump by flow diverter.Oral digestion chamber 2 left end outlets are by diverting valve, and connection valve is connected with oral digestion product ware 16 with constant flow pump.The middle part of oral digestion chamber 2 left end outlets is connected with peptic digest chamber 3 left end sample feeding pipes with connection valve by emptying delivery pipe.
Peptic digest district B: 3 inside, the straight-through peptic digest chamber of peptic digest chamber 3 left end sample feeding pipes.Peptic digest chamber 3 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with gastric juice ware 8 with a constant flow pump by flow diverter.Peptic digest chamber 3 right-hand member outlets are connected with a conduit by diverting valve, and this conduit other end is connected with peptic digest product ware 17 with constant flow pump by connection valve.Also be connected with small intestine digestion chamber 4 right-hand member sample feeding pipes by emptying delivery pipe in diverting valve junction.
Small intestine digestion zone C: 4 inside, the straight-through small intestine digestion chamber of small intestine digestion chamber 4 right-hand member sample feeding pipes.Small intestine digestion chamber 4 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with intestinal fluid ware 9 with a constant flow pump by flow diverter.Small intestine digestion chamber 4 left end outlets are connected with a conduit by diverting valve, and this conduit other end is connected with small intestine digestion product ware 18 with constant flow pump by connection valve.Also be connected with large intestine leading portion glycolysis chamber 5 left end sample feeding pipes by emptying delivery pipe in diverting valve junction.
Large intestine leading portion glycolysis district D: 5 inside, the straight-through large intestine leading portion glycolysis chamber of large intestine leading portion glycolysis chamber 5 left end sample feeding pipes.Large intestine leading portion glycolysis chamber 5 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with large intestine leading portion glycolysis nutrient culture media ware 10 with a constant flow pump by flow diverter.Large intestine leading portion glycolysis chamber 5 right-hand member outlets are connected with one end of airtight the first sieves 14 by diverting valve and conduit, first airtight sieves 14 other ends are connected with one end of the first dialysate chamber 12 by conduit, and first dialysate chamber 12 other ends are by a constant flow pump and large intestine leading portion glycolysis product anaerobism ware 19.Also be connected with large intestine back segment glycolysis chamber 6 right-hand member sample feeding pipes by emptying delivery pipe in diverting valve junction.
Large intestine back segment glycolysis district E: 6 inside, the straight-through large intestine back segment glycolysis chamber of large intestine back segment glycolysis chamber 6 left end sample feeding pipes.Large intestine back segment glycolysis chamber 6 is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously.Junction at two locellus is connected with large intestine back segment glycolysis nutrient culture media ware 11 with a constant flow pump by flow diverter.Large intestine back segment glycolysis chamber 5 left end outlets are connected with second airtight sieves 15 one end with conduit by diverting valve, second airtight sieves 15 other ends are connected with second dialysate chamber 13 one end by conduit, and second dialysate chamber 13 other ends are by a constant flow pump and large intestine back segment glycolysis product anaerobism ware 20.Also reclaiming ware 21 by emptying delivery pipe with residue glycolysis product in diverting valve junction is connected.
Using method: soy bean proteinous soln is made into certain concentration solution (5 mg/ml).To shift to an earlier date ready saliva, gastric juice, intestinal fluid, large intestine leading portion glycolysis nutrient culture media, large intestine back segment glycolysis nutrient culture media joins respectively in saliva ware 7, gastric juice ware 8, intestinal fluid ware 9, large intestine leading portion glycolysis nutrient culture media ware 10, large intestine back segment glycolysis nutrient culture media ware 11.Open whole water bath with thermostatic control systems and keep 37 DEG C of constant temperature.Open whole constant flow pumps and diverting valve.All the speed of constant flow pump is controlled at 5 ml/min.Soy bean proteinous soln is added from the sample inlet 1 of the top of digestion glycolysis system and device, saliva joins oral digestion chamber 2 by constant flow pump and flow diverter from saliva ware 7, carry out oral digestion simulation, after oral digestion 15 min, the postdigestive collection of products of a part obtains soybean protein oral digestion product in oral digestion product ware 16, and another part digestion product enters in the peptic digest chamber 3 in peptic digest district B with emptying delivery pipe.Gastric juice joins peptic digest chamber 3 by constant flow pump and flow diverter from gastric juice ware 8, carry out peptic digest simulation, after peptic digest 5 h, the postdigestive collection of products of a part obtains soybean protein peptic digest product in peptic digest product ware 17, and another part digestion product enters in the small intestine digestion chamber 4 in the C of small intestine digestion zone with emptying delivery pipe.Intestinal fluid joins small intestine digestion chamber 4 by constant flow pump and flow diverter from intestinal fluid ware 9, small intestine digests after 5 h, the postdigestive collection of products of a part obtains soybean protein small intestine digestion product in small intestine digestion product ware 18, and another part digestion product enters in the large intestine leading portion glycolysis chamber 5 in large intestine leading portion glycolysis district D with emptying delivery pipe.Large intestine leading portion glycolysis nutrient culture media joins large intestine leading portion glycolysis chamber 5 by constant flow pump and flow diverter from large intestine leading portion glycolysis nutrient culture media ware 10, after large intestine leading portion glycolysis 12 h, product after part glycolysis is collected in large intestine leading portion glycolysis product anaerobism ware 19 and is obtained soybean protein large intestine leading portion glycolysis product by the first airtight sieves 14 and the first dialysate chamber 12, and another part glycolysis product enters in the large intestine back segment glycolysis chamber 6 in large intestine back segment glycolysis district E with emptying delivery pipe.Large intestine back segment glycolysis nutrient culture media joins large intestine back segment glycolysis chamber 6 by constant flow pump and flow diverter from large intestine back segment glycolysis nutrient culture media ware 11, after large intestine back segment glycolysis 12 h, product after part glycolysis is collected in large intestine back segment glycolysis product anaerobism ware 20 and is obtained soybean protein large intestine back segment glycolysis product by the second airtight sieves 15 and the second dialysate chamber 13, and another part glycolysis product enters residue glycolysis product with emptying delivery pipe 15 and reclaims in ware 21.Simulation digestion glycolysis process repeats 3 times.
Oral digestion product ware 16 is placed in 5 minutes activity with removal ptyalin of boiling water bath (preparing in advance), detects afterwards the oral digestion situation (the change of molecular weight) of soybean protein in oral digestion product ware 16 with liquid phase method.Peptic digest product ware 17 is placed in boiling water bath (in advance prepare) 5 minutes to remove pepsic activity, detects afterwards the peptic digest situation (the change of molecular weight) of soybean protein in peptic digest product ware 17 with liquid phase method.Small intestine digestion product ware 18 is placed in boiling water bath (preparing in advance) and removes pancreatin (comprising amylopsin and pancreatic lipase) and tryptic activity to remove in 5 minutes, detects afterwards the small intestine digestion situation (the change of molecular weight) of soybean protein in small intestine digestion product ware 18 with liquid phase method.Large intestine leading portion glycolysis product anaerobism ware 19 (is prepared in advance) 30 minutes to remove the activity of residue flora in boiling water bath, and detects the large intestine leading portion glycolysis product of soybean protein in large intestine leading portion glycolysis product anaerobism ware 19 with vapor phase method.Large intestine back segment glycolysis product anaerobism ware 20 (is prepared in advance) 30 minutes to remove the activity of residue flora in boiling water bath, and detects the large intestine back segment glycolysis product of soybean protein in large intestine back segment glycolysis product anaerobism ware 20 with vapor phase method.
Testing result: this soybean protein can not be decomposed in the simulation digestion of oral cavity.After stomach simulation digestion, molecular weight significantly reduces, and shows that this soybean protein is significantly decomposed in human stomach's digestion.After small intestine simulation digestion, molecular weight significantly reduces, and shows that this soybean protein is decomposed in a large number in human small intestine's digestion.This soybean protein can produce a certain amount of SCFA after the glycolysis of large intestine leading portion and back segment glycolysis, and can increase acetic acid, the amount of propionic acid and butyric acid.

Claims (2)

1. simulate the device of human body digestion glycolysis system for one kind, it is characterized in that comprising sample inlet (1), oral digestion chamber (2), peptic digest chamber (3), small intestine digestion chamber (4), large intestine leading portion glycolysis chamber (5), large intestine back segment glycolysis chamber (6), saliva ware (7), gastric juice ware (8), intestinal fluid ware (9), large intestine leading portion glycolysis nutrient culture media ware (10), large intestine back segment glycolysis nutrient culture media ware (11), the first dialysate chamber (12), the second dialysate chamber (13), airtight the first sieves (14), airtight the second sieves (15), oral digestion product ware (16), peptic digest product ware (17), small intestine digestion product ware (18), large intestine leading portion glycolysis product anaerobism ware (19), large intestine back segment glycolysis product anaerobism ware (20), residue glycolysis product reclaims ware (21),
Sample inlet (1) is connected with oral digestion chamber (2) right-hand member sample feeding pipe with diverting valve by connection valve, inside, the straight-through oral cavity digestion chamber (2) of oral digestion chamber (2) right-hand member sample feeding pipe; Oral digestion chamber (2) are all equipped with the water bath with thermostatic control circulation system in outside simultaneously; Oral digestion chamber (2) left end is connected with saliva ware (7) with a constant flow pump by flow diverter; Oral digestion chamber (2) left end outlet is by diverting valve, and connection valve is connected with oral digestion product ware (16) with constant flow pump; The middle part of oral digestion chamber (2) left end outlet is connected with peptic digest chamber (3) left end sample feeding pipe with connection valve by emptying delivery pipe;
Inside, the straight-through peptic digest chamber (3) of peptic digest chamber (3) left end sample feeding pipe; Peptic digest chamber (3) is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously; Junction at two locellus is connected with gastric juice ware (8) with a constant flow pump by flow diverter; Peptic digest chamber (3) right-hand member outlet is connected with a conduit by diverting valve, and this conduit other end is connected with peptic digest product ware (17) with constant flow pump by connection valve; Also be connected with small intestine digestion chamber (4) right-hand member sample feeding pipe by emptying delivery pipe in diverting valve junction;
Inside, the straight-through small intestine digestion chamber (4) of small intestine digestion chamber (4) right-hand member sample feeding pipe; Small intestine digestion chamber (4) is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously; Junction at two locellus is connected with intestinal fluid ware (9) with a constant flow pump by flow diverter; Small intestine digestion chamber (4) left end outlet is connected with a conduit by diverting valve, and this conduit other end is connected with small intestine digestion product ware (18) with constant flow pump by connection valve; Also be connected with large intestine leading portion glycolysis chamber (5) left end sample feeding pipe by emptying delivery pipe in diverting valve junction;
Inside, the straight-through large intestine leading portion glycolysis chamber (5) of large intestine leading portion glycolysis chamber (5) left end sample feeding pipe; Large intestine leading portion glycolysis chamber (5) is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously; Junction at two locellus is connected with large intestine leading portion glycolysis nutrient culture media ware (10) with a constant flow pump by flow diverter; Large intestine leading portion glycolysis chamber (5) right-hand member outlet is connected with one end of airtight the first sieves (14) by diverting valve and conduit, airtight the first sieves (14) other end is connected with one end of the first dialysate chamber (12) by conduit, and the first dialysate chamber (12) other end is by a constant flow pump and large intestine leading portion glycolysis product anaerobism ware (19); Also be connected with large intestine back segment glycolysis chamber (6) right-hand member sample feeding pipe by emptying delivery pipe in diverting valve junction;
Inside, the straight-through large intestine back segment glycolysis chamber (6) of large intestine back segment glycolysis chamber (6) left end sample feeding pipe; Large intestine back segment glycolysis chamber (6) is divided into two locellus, and two locellus are all equipped with the water bath with thermostatic control circulation system in outside, and two locellus are fixedly connected with by two standing valves simultaneously; Junction at two locellus is connected with large intestine back segment glycolysis nutrient culture media ware (11) with a constant flow pump by flow diverter; Large intestine back segment glycolysis chamber (5) left end outlet is connected with airtight the second sieves (15) one end with conduit by diverting valve, airtight the second sieves (15) other end is connected with the second dialysate chamber (13) one end by conduit, and the second dialysate chamber (13) other end is by a constant flow pump and large intestine back segment glycolysis product anaerobism ware (20); Also reclaiming ware (21) by emptying delivery pipe with residue glycolysis product in diverting valve junction is connected.
2. the using method of the device of simulation human body digestion glycolysis system claimed in claim 1, is characterized in that:
Material to be digested is dissolved or dispersed in distilled water and makes determinand solution or dispersion liquid with finite concentration, will shift to an earlier date ready saliva, gastric juice, intestinal fluid, large intestine leading portion glycolysis nutrient culture media, large intestine back segment glycolysis nutrient culture media and join respectively in saliva ware (7), gastric juice ware (8), intestinal fluid ware (9), large intestine leading portion glycolysis nutrient culture media ware (10), large intestine back segment glycolysis nutrient culture media ware (11); Open whole water bath with thermostatic control systems and keep 37 DEG C of constant temperature; Open whole constant flow pumps and diverting valve; All the speed of constant flow pump is controlled at 4-6 ml/min; Determinand solution or dispersion liquid are added from the sample inlet (1) of the top of digestion glycolysis system and device, saliva joins oral digestion chamber (2) by constant flow pump and flow diverter from saliva ware (7), carry out oral digestion simulation, after oral digestion 10-15 min, the postdigestive collection of products of a part is in oral digestion product ware (16), and another part digestion product enters in the peptic digest chamber (3) in peptic digest district (B) with emptying delivery pipe; Gastric juice joins peptic digest chamber (3) by constant flow pump and flow diverter from gastric juice ware (8), carry out peptic digest simulation, after peptic digest 5-6 h, the postdigestive collection of products of a part is in peptic digest product ware (17), and another part digestion product enters in the small intestine digestion chamber (4) in small intestine digestion zone (C) with emptying delivery pipe; Intestinal fluid joins small intestine digestion chamber (4) by constant flow pump and flow diverter from intestinal fluid ware (9), after small intestine digestion 5-6 h, the postdigestive collection of products of a part is in small intestine digestion product ware (18), and another part digestion product enters in the large intestine leading portion glycolysis chamber (5) in large intestine leading portion glycolysis district (D) with emptying delivery pipe; Large intestine leading portion glycolysis nutrient culture media joins large intestine leading portion glycolysis chamber (5) by constant flow pump and flow diverter from large intestine leading portion glycolysis nutrient culture media ware (10), after large intestine leading portion glycolysis 10-12 h, product after part glycolysis is collected in large intestine leading portion glycolysis product anaerobism ware (19) by airtight the first sieves (14) and the first dialysate chamber (12), and another part glycolysis product enters in the large intestine back segment glycolysis chamber (6) in large intestine back segment glycolysis district (E) with emptying delivery pipe; Large intestine back segment glycolysis nutrient culture media joins large intestine back segment glycolysis chamber (6) by constant flow pump and flow diverter from large intestine back segment glycolysis nutrient culture media ware (11), after large intestine back segment glycolysis 10-12 h, product after part glycolysis is collected in large intestine back segment glycolysis product anaerobism ware (20) by airtight the second sieves (15) and the second dialysate chamber (13), and another part glycolysis product enters residue glycolysis product with emptying delivery pipe and reclaims in ware (21); Simulation digestion glycolysis process repeats 3 times;
Oral digestion product ware (16) is placed in boiling water bath (preparing in advance) a few minutes to remove the activity of ptyalin, detects afterwards the oral digestion situation of determinand in oral digestion product ware (16) with liquid phase method; Peptic digest product ware (17) is placed in boiling water bath (preparing in advance) a few minutes to remove pepsic activity, detects afterwards the peptic digest situation of determinand in peptic digest product ware (17) with liquid phase method; Small intestine digestion product ware (18) is placed in boiling water bath (preparing in advance) a few minutes to remove except pancreatin (comprising amylopsin and pancreatic lipase) and tryptic activity, detects afterwards the small intestine digestion situation of determinand in small intestine digestion product ware (18) with liquid phase method; Large intestine leading portion glycolysis product anaerobism ware (19) in boiling water bath (in advance prepare) 30-40 minute to remove activity of residue flora, and detect large intestine leading portion glycolysis product in large intestine leading portion glycolysis product anaerobism ware (19) with vapor phase method; Large intestine back segment glycolysis product anaerobism ware (20) in boiling water bath (in advance prepare) 30-40 minute to remove activity of residue flora, and detect large intestine back segment glycolysis product in large intestine back segment glycolysis product anaerobism ware (20) with vapor phase method.
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